Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Journal of Clinical Rehabilitative Tissue Engineering Research》 2009-28
Add to Favorite Get Latest Update

Package of recombinant adeno-associated virus encoding cell division cycle 2-siRNA

Wei Jia-jun,Wang Zhuo-ran Department of Neurology,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei Province,China  
BACKGROUND:Cell division cycle 2 (cdc2) plays an important role in the course of neuronal degeneration in neurodegenerative diseases such as Alzheimer's disease. The silencing of cdc2 gene with adeno-associated virus vector might protect the neurons in neurodegenerative diseases. OBJECTIVE:To pack recombinant adeno-associated virus (rAAV) encoding cdc2-siRNA. DESIGN,TIME AND SETTING:Blank control experiment was performed in the laboratory of Department of Neurology at Wuhan Tongji Hospital between October 2007 and August 2008. MATERIALS:Helper Free adeno-associated virus system (pAAV-MCS-EGFP,p-RC,p-Helper) and AAV-293 cells were purchased from Stratagene. METHODS:The pAAV-MCS-U6-cdc2-siRNA-EGFP expressing plasmid was constructed by molecular biological techniques. The cotransfection of this plasmid,together with p-RC and p-Helper into AAV-293 cells was mediated by calcium acid phosphate. The rAAV encoding cdc2-siRNA (rAAV-U6-cdc2-siRNA-EGFP) was packed and harvested. The silencing effect of this virus on cdc2 gene expression in AAV-293 cells was assessed by Western Blot,and its titer was determined by spot hybridization method. MAIN OUTCOME MEASURES:sequencing of pAAV-MCS-U6-cdc2-siRNA-EGFP plasmid inserting U6-cdc2-siRNA; 3 plasmid pAAV-MCS-U6-cdc2-siRNA-EGFP,p-RC,p-Helper cotransfecting AAV-293 cells; cdc2 expression after rAAV-U6-cdc2-siRNA-EGFP infected AAV-293 cells. RESULTS:① The successful construction of U6-cdc2-siRNA in pAAV-MCS-EGFP plasmid was proved by DNA sequencing. ② Green fluorescent protein was expressed in AAV-293 cells,and the contransfection was working well. ③ The cdc2 gene expression in AAV-293 cells was down-regulated markedly after infection of rAAV-U6-cdc2-siRNA-EGFP. ④ The titer of rAAV-U6-cdc2-siRNA-EGFP was 1x1012 v.g/mL. CONCLUSION:The package of rAAV encoding cdc2-siRNA is successful. It can silence cdc2 gene effectively.
【CateGory Index】: R346
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 SAI Jia-ming,ZHANG Hui-qin(Orthopaedic Laboratory,Qingdao Hospital of Orthopaedics and Traumatology,Qingdao 266021,China);Biological effects of co-transfection of TGFβ_1 and VEGF mediated by AAVon promoting healing of the cutaneous ulcer in diabetes mellitus[J];Journal of Traumatic Surgery;2007-05
2 HU Bin,YANG Yan,LIU Jia,et al. Institute of Clinical Immunology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Inhibition of HbsAg and HbeAg expression of recombinant adeno-associated virus encoding HBsAg-shRNA in HepG2.215 cells in vitro[J];Journal of Clinical Hepatology;2010-03
3 Sai Jiaming,Hu Yougu,Wang Dechun.Department of Orthopedics,Hospital Affiliated Qingdao Medical Colledge,Qingdao,266021;Construction of AAV-TGFβ_1 and Comparision of Its Biological Effects on Proteoglycan Synthesis of Nucleus Pulpous Cells with AV-TGFβ_1[J];Chinese Journal of Bone and Joint Injury;2008-02
4 JIANG Feng,WANG Xue-zhen,BU Bi-tao,ZHANG Su-ming,ZHANG Min.Department of Neurology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Construction and Identification of Lentivector-mediated CDK4-RNAi[J];Neural Injury and Functional Reconstruction;2008-06
5 DU Shu-min,CAO Yi-cheng,LI Xin-jian (School of Bioscience and Bioengineering, South China University of Technology, Guangzhou 510006, China);Construction of Recombinat Adeno-assosiated Virus Vector Containing a Fusion Rspo1-EGFP Gene[J];Modern Food Science and Technology;2009-09
6 Chen Dejie, Tan Zui, Xie Youli, et al Department of Vascular Surgery, Zhongnan Hospital, Wuhan University, Wuhan 430071,China;Construction of Adeno-Associated Virus Co-expression System for Human Angiopoietin 1 and VEGF Gene[J];Journal of Hubei Medical University;2004-02
7 ZHAO Xiao-yan1,2,HU Shen-jiang2(1.Department of Cardiology,the Fifth People's Hospital of Zhengzhou,Zhengzhou Henan,450003;(2.Institute of Cardiology,the First Affiliated Hospital,College of Medicine,Zhejiang University,Hangzhou,Zhejiang 310003 China);Angiogenin Gene Transfer Improves Left Ventricular Function in Rats with Myocardial Infarction[J];Journal of Henan Medical College for Staff and Workers;2007-03
8 SAI JiaMing, HU YouGu & WANG DeChun Department of Orthopedic Surgery, Affiliated Hospital of Medical College, Qingdao University, Qingdao 266003, China Orthopae-dic Laboratory, The Orthopaedics and Traumatology Hospital, Qingdao 266003, China;Effects of adeno-associated virus (AAV) of transforming growth factors β_1 and β_3 (TGFβ_(1,3)) on promoting synthesis of glycosaminoglycan and collagen type II of dedifferentiated nucleus pulposus (NP) cells[J];中国科学(C辑:生命科学)(英文版);2007-05
9 GAO Ping jin , ZHU Ding liang (Shanghai Institue of Hypertension, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025,China);Gene therapy for essential hypertension[J];Basic Medical Sciences and Clinics;2002-06
10 CHEN Li, CHEN Haoming, LI Ge, LU Daru & XUE Jinglun State Key Laboratory of Genetic Engineering, Institute of Genetics,School of Life Sciences, Fudan University, Shanghai 200433, China Correspondence should be addressed to Xue Jinglun (e-mail: jlxue@ fudan.ac.cn);Novel rAAV production system with low contamina-tion of helper virus[J];科学通报(英文版);2003-05
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved